Figures and Methods for : Cullin 5 regulates Dab 1 protein levels and neuron positioning during cortical development

Supplementary Methods Plasmids and Constructs pCMXGFP-Dab1, pCMXGFP-Dab1, pCMXGFP-Dab1, pCMXCFP-Dab1, pCMXGFP-Dab1 and pCMXGFP-Dab1 expression plasmids were constructed by cutting out fragments containing the corresponding mutants from pBSK HADab1 (Howell et al., 2000) with EcoRI/XbaI and subcloning into pCMXGFP-p1k1 with EcoRI/NheI. The pCAG-Dab1GFP was described before (Xu et al., 2005). Different phosphorylation site mutants were generated by swapping in BstXI /BglII fragments from the corresponding pCMXGFP-Dab1 mutants. The following constructs were kind gifts from the following individuals: pME18S-Fyn(B) WT and KM(kinase dead), from Tohru Tezuka; pEF-FlagmSOCS1/3, Neil Nathanson; pEF-Flag-mSOCS2, Chris Greenhalgh (Nicholson et al., 1999); pCEFL-hCbl-b-HA and corresponding Cbl, Cbl-3 and myc-E6AP constructs, Stanley Lipkowitz (Ettenberg et al., 1999; Hatakeyama et al., 1997; Keane et al., 1999); pCMX Flag-mLNX, Jane McGlade (Nie et al., 2002); Rnf5, Ze'ev Ronai (Didier et al., 2003); and Hakai, Yasu Fujita (Fujita et al., 2002).